Ginkgo biloba is grown worldwide as an ornamental plant for its golden leaf color. However, the regulatory mechanism of leaf coloration in G. biloba remains unclear. Here, we compared G. biloba gold-colored mutant leaves and normal green leaves in cytological, physiological and transcriptomic terms. We found that chloroplasts of the mutant were fewer and smaller, and exhibited ruptured thylakoid membranes, indistinct stromal lamellae and irregularly arranged vesicles. Physiological experiments also showed that the mutant had a lower chlorophyll, lower flavonoid and higher carotenoid contents (especially lutein). We further used transcriptomic sequencing to identify 116 differentially expressed genes (DEGs) and 46 transcription factors (TFs) involved in chloroplast development, chlorophyll metabolism, pigment biosynthesis and photosynthesis. Among these, the chlorophyll biosynthesis-related PPO showed down-regulation, while chlorophyll degradation-related NYC/NOL had up-regulated expression in mutant leaves. Z-ISO, ZDS, and LCYE, which are involved in carotenoid biosynthesis were up-regulated. Quantitative real-time PCR (RT-qPCR) further confirmed the altered expression levels of these genes at three stages. The alteration of PPO and NYC/NOL gene expression might affect chlorophyll biosynthesis and promote degradation of chlorophyll b to chlorophyll a, while the up-regulated genes Z-ISO, ZDS and LCYE enhanced carotenoid accumulation. Consequently, changes in the ratio of carotenoids to chlorophylls were the main factors driving the golden leaf coloration in the mutant G. biloba.
To gain insights into the regulatory networks of miRNAs related to golden colour formation in Ginkgo biloba leaves, we constructed an sRNA library of golden-green striped mutant leaves. A total of 213 known miRNAs comprising 54 miRNA families were obtained, and 214 novel miRNAs were identified in the mutant leaves. We further constructed a normal green leaf sRNA library as a control and compared the expression of miRNAs between mutant and normal leaves. We found 42 known and 54 novel differential expression candidate miRNAs; 39 were up-regulated and 57 down-regulated in mutants compared to normal leaves. Our transcriptome analysis and annotation of the predicted targets indicated that the potential roles of miRNAs in G. biloba leaves included involvement in the ‘Glutathione metabolism’, ‘Plant circadian rhythm’, and ‘Phenylalanine metabolism’ categories. miRNAs and their targets were further validated by qRT-PCR. The expression of miR159a and miR159c, in particular, was significantly higher in mutant leaves than in normal leaves, while their potential target gene CLT3, which is associated with chloroplast development, displayed the opposite expression pattern. In addition, the expression of miR396g-3p and miR396h was also significantly higher in mutant leaves than in normal leaves, while the target genes ABP1 (auxin-related gene) and PPR32 (chloroplast RNA editing protein), respectively, showed the opposite expression pattern. Combined with the transcriptome analysis, these data suggest that miR159, miR396, and their targets may participate in chloroplast development and hormone metabolism to regulate colour formation in G. biloba leaves.
Manufacturing agglomeration promotes rapid economic development while also causing severe environmental pollution. This paper investigates the impact and mechanism of manufacturing agglomeration on haze pollution from the Chinese city level. Furthermore, we discuss the moderating effect and threshold effect of the three urban forms of urban external shape complexity, urban compactness, and urban fragmentation on the relationship between the two. The result shows: (1) The aggregation of the manufacturing industry presents an inverted U-shaped characteristic of promoting first and then inhibiting haze pollution in China's overall, eastern and central regions. (2) The complexity of the city's external shape and the city's fragmentation has a positive moderating effect on the relationship between manufacturing agglomeration and haze pollution. And urban fragmentation shows a negative moderating effect on the relationship between the two when the level of manufacturing agglomeration is on the right side of the inverted U-shaped curve. (3) The urban form shows a significant double threshold characteristic for haze pollution, increasing the complexity of the city's external shape and the city's fragmentation. The agglomeration of manufacturing shows the characteristics of first inhibiting and then promoting haze pollution. As urban compactness increases, the inhibitory effect of manufacturing agglomeration on haze pollution increases.
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